Organization and structure of an Onchocerca ß-tubulin gene

RESUMO

In filarial worms, as in other eukaryotes, microtubules are essential multifunctional components. The major protein of microtubules is tubulin, a heterodimer of two distinct polypeptides, Ó e ß.Tubulin is particulary important in helminthic parasites as a target for anthelminthic benzimidazoles, wich bind to it and inhibit microtubule assembly. Two genomic Onchocerca gibsoni libraries were constructed in NM1149(EcoRi and HindIII). Three clones accounted for the entire gene one from the EcoRi library (using a Plasmodium falciparum probe) containing the central part of the gene, and two from the HindIII library (using as probes PCR amplified fragments from the ends of the EcoRI clone) which, respectively, contained the 5'- and 3' -ends of the gene. The sequencing procedure for the EcoRI clone relied on the construction of a double-digested DraI/HindIII shotgun library. A number of recombinants were sequenced and aligned with each other for comparison. The sequencing of the overlapping 5' - and 3'-end clones was done by ...